目的 探究止嗽化痰丸（ZSHT）对哮喘小鼠的作用效果，并基于代谢组学解析其作用机制。方法 SPF级雄性BALB/c小鼠适应性喂养1周后随机分为对照组、模型组、阳性药孟鲁司特钠(MS,1.30 mg·kg^-1)组与咳喘宁(KCN,470 mg·kg^-1)组和ZSHT低、中、高剂量(300、600、1 190 mg·kg^-1)组。在第0、7、14天，除对照组外其余小鼠ip卵清白蛋白（OVA）致敏，连续雾化激发7 d，每次激发前30 min对各给药组小鼠ig给予相应药物进行治疗。通过评价小鼠肺功能、观察肺组织病理变化评价ZSHT对哮喘小鼠的药效；酶联免疫吸附测定（ELISA）小鼠血清中炎症因子水平；流式细胞术检测小鼠肺组织中活性氧、线粒体膜电位、凋亡水平以及外周血中免疫细胞比例；基于超高效液相色谱-质谱联用技术（UPLC-MS/MS）对不同处理组小鼠的血清样本进行代谢组学分析，筛选差异代谢物并富集相关代谢通路。结果 与模型组比较，ZSHT 3种剂量均可以抑制哮喘小鼠气道高反应性，低剂量显著增加潮气量，降低气道狭窄指数、最大呼气流速、呼出50%气量时对应的呼气流速(EF₅₀)，减少咳喘次数，延长咳喘潜伏期（P<0.05、0.01）；显著缓解哮喘小鼠肺部炎症以及胶原纤维沉积（P<0.01），当给药剂量增至中、高剂量时，其治疗效果未随剂量增加而提升。ELISA结果同样显示，ZSHT低剂量可显著升高血清中γ干扰素（IFN-γ）水平，显著降低血清中的免疫球蛋白E（IgE）、白细胞介素4（IL-4）水平（P<0.01），同时可以显著缓解哮喘小鼠肺组织中氧化应激和凋亡状态（P<0.01），改善外周血免疫细胞Th1/Th2比例失衡，升高Treg细胞水平（P<0.01）。基于代谢组学进一步探究其作用机制，筛选出模型组与ZSHT低剂量组差异代谢物62个，其中标志性差异代谢物13个，主要富集在花生四烯酸代谢、苯丙氨酸代谢、组氨酸代谢、视黄醇代谢、半乳糖代谢5个代谢通路。结论 ZSHT对OVA诱导的哮喘模型小鼠有明确干预作用，可通过调节Th1/Th2免疫平衡、降低炎症因子水平，影响花生四烯酸、间香豆素酸等关键差异代谢物的生成与转化，进而调控与炎症相关的核心通路，改善小鼠病理状态。

Objective To investigate the effect of Zhisou Huatan Pills（ZSHT） on asthmatic mice and to explore its mechanism of action based on metabolomics, providing experimental evidence for the clinical application of ZSHT in the treatment of asthma. Methods SPF-grade male BALB/c mice were adaptively fed for one week and then randomly divided into the control group, model group, positive drug montelukast sodium(MS, 1.30 mg·kg^-1) group, Kechuanning(KCN, 470 mg·kg^-1) group, and ZSHT low-, medium-, and high-dose(300, 600, 1 190 mg·kg^-1) groups. On days 0, 7, and 14, except for the control group, the remaining mice were sensitized intraperitoneally with ovalbumin（OVA）, and then continuously nebulized for seven days. Thirty minutes before each nebulization, the mice in each drug administration group were treated ig with the corresponding drugs. The efficacy of ZSHT on asthmatic mice was evaluated by assessing lung function and observing pathological changes in lung tissue. The levels of inflammatory factors in mouse serum were determined by enzyme-linked immunosorbent assay（ELISA）. The levels of reactive oxygen species, mitochondrial membrane potential, apoptosis in lung tissue and the proportion of immune cells in peripheral blood were detected by flow cytometry. Based on ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）, the serum samples of different treatment groups were subjected to metabolomics analysis to screen for differential metabolites and enrich related metabolic pathways. On days 0, 7, and 14, except for the control group, the remaining mice were sensitized by intraperitoneal injection of ovalbumin（OVA）, and then continuously challenged by nebulization for seven days. Thirty minutes before each challenge, the mice in the treatment groups were given the corresponding drugs by gavage. The efficacy of ZSHT on asthmatic mice was evaluated by assessing lung function and observing pathological changes in lung tissue. The levels of inflammatory factors in mouse serum were determined by ELISA. The levels of reactive oxygen species, mitochondrial membrane potential, apoptosis in lung tissue and the proportion of immune cells in peripheral blood were detected by flow cytometry. Metabolomics analysis was performed on serum samples from different treatment groups using ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）, and differential metabolites were screened and related metabolic pathways were enriched. Results Compared with the model group, all three doses of ZSHT could inhibit airway hyperresponsiveness in asthmatic mice. The low dose significantly increased tidal volume, reduced airway narrowing index, maximum expiratory flow rate, and expiratory flow rate at 50% of the exhaled volume(EF₅₀), reduced the frequency of coughing and wheezing, and prolonged the latency of coughing and wheezing（P ＜0.05, 0.01）. It significantly alleviated lung inflammation and collagen fiber deposition in asthmatic mice（P ＜0.01）. When the dosage was increased to medium and high doses, the therapeutic effect did not increase with the dose. ELISA results also showed that the low dose of ZSHT could significantly increase the level of interferon-γ（IFN-γ） in serum, significantly reduce the levels of immunoglobulin E（IgE） and interleukin-4（IL-4） in serum（P ＜0.01）, and significantly alleviate oxidative stress and apoptosis in lung tissue of asthmatic mice（P ＜0.01）, improve the imbalance of Th1/Th2 ratio in peripheral blood immune cells, and increase the level of Treg cells（P ＜0.01）. Further exploration of its mechanism of action based on metabolomics identified 62 differential metabolites between the model group and the low-dose ZSHT group, among which 13 were signature differential metabolites, mainly enriched in arachidonic acid metabolism, phenylalanine metabolism, histidine metabolism, retinol metabolism, and galactose metabolism pathways. Conclusion ZSHT has a definite intervention effect on the asthma model mice induced by OVA, with the low dose being the most effective. It can regulate the Th1/Th2 immune balance, reduce inflammatory factors, and affect the generation and transformation of key differential metabolites such as arachidonic acid and coumaric acid, thereby regulating the core pathways related to inflammation and improving the pathological state of mice.

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国家重点研发计划资助项目(2017YFC1702800,2019YFC1708802); 止嗽化痰丸(加减马兜铃)药效学研究资助项目(14208126)